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Automatic Evaluation of Collagen Fiber Directions from Polarized Light Microscopy Images

Published online by Cambridge University Press:  08 May 2015

Kamil Novak*
Affiliation:
Institute of Solid Mechanics, Mechatronics and Biomechanics, Brno University of Technology, Czech Republic
Stanislav Polzer
Affiliation:
Institute of Solid Mechanics, Mechatronics and Biomechanics, Brno University of Technology, Czech Republic
Michal Tichy
Affiliation:
2nd Department of Pathology and Anatomy, St. Anne’s University Hospital, Czech Republic
Jiri Bursa
Affiliation:
Institute of Solid Mechanics, Mechatronics and Biomechanics, Brno University of Technology, Czech Republic
*
*Corresponding author. kamil_novak@centrum.cz
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Abstract

Mechanical properties of the arterial wall depend largely on orientation and density of collagen fiber bundles. Several methods have been developed for observation of collagen orientation and density; the most frequently applied collagen-specific manual approach is based on polarized light (PL). However, it is very time consuming and the results are operator dependent. We have proposed a new automated method for evaluation of collagen fiber direction from two-dimensional polarized light microscopy images (2D PLM). The algorithm has been verified against artificial images and validated against manual measurements. Finally the collagen content has been estimated. The proposed algorithm was capable of estimating orientation of some 35 k points in 15 min when applied to aortic tissue and over 500 k points in 35 min for Achilles tendon. The average angular disagreement between each operator and the algorithm was −9.3±8.6° and −3.8±8.6° in the case of aortic tissue and −1.6±6.4° and 2.6±7.8° for Achilles tendon. Estimated mean collagen content was 30.3±5.8% and 94.3±2.7% for aortic media and Achilles tendon, respectively. The proposed automated approach is operator independent and several orders faster than manual measurements and therefore has the potential to replace manual measurements of collagen orientation via PLM.

Type
Biological Applications and Techniques
Copyright
© Microscopy Society of America 2015 

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